Single-use manifolds for automated, aseptic handling of solutions in bioprocessing applications

ABSTRACT

Presteralized manifolds are provided which are designed for sterile packaging and single-use approaches. Disposable tubing and flexible-wall containers are assembled via aseptic connectors. These manifolds are adapted to interact with other equipment which can be operated by a controller which provides automated and accurate delivery of biotechnology fluid. The manifold also can be used in conjunction with one or more sensors such as pressure and conductivity sensors that interact with the controller or are connected to a separate user interface. An aseptic environment obtains avoiding or reducing cleaning and quality assurance procedures.

CROSS REFERENCE TO RELATED APPLICATION

This is a continuation of application Ser. No. 11/294,297, filed Dec. 5,2005, which is a continuation-in-part of application Ser. No.10/764,624, filed Jan. 26, 2004, now U.S. Pat. No. 7,052,603, which is adivisional of application Ser. No. 10/172,082 filed Jun. 14, 2002, nowU.S. Pat. No. 6,712,963, each incorporated hereinto by reference.

FIELD OF THE INVENTION

The invention generally relates to the aseptic transfer of solutions outof one or more biological fluid and/or process fluid storage or supplycontainers. Single-use manifold systems carry out transfers needed inbioprocessing applications. With the invention, automated dispensing isaccomplished, preferably in association with one or more disposableconductivity sensors and often with one or more remotely controlledpinch valves.

BACKGROUND OF THE INVENTION

Good manufacturing practices and governmental regulations are at thecore of any pharmaceutical, biotechnology and bio-medical manufacturingprocess or procedure. Such manufacturing processes and procedures aswell as associated equipment must undergo mandated, often lengthy andcostly validation procedures. Similar issues exist for sensors whenneeded in such systems, such as conductivity sensors.

For example, the equipment used for the separation and purification ofbiomedical products must, for obvious reasons, meet stringentcleanliness requirements. The cleaning validation of new orre-commissioned purification equipment (including sensor equipment suchas equipment for preparative chromatography or tangential flowfiltration) may require as many as 50 test-swabs of exposed surfaces andsubsequent biological assays of such test-swabs. For a single piece ofpurification equipment, the associated and reoccurring cost of a singlecleaning validation may readily exceed multiple thousands of dollars.

To reduce such cleaning validation costs and expenses, and/or to reducethe occasions when cleaning is needed or required, the pharmaceuticaland biotech industries are increasingly employing, pre-sterilized,single-use, plastic tubing and collapsible, plastic bags for solutiontransfer and storage. Sterilization is accomplished by exposing thecomplete tube/bag manifold to gamma irradiation, or to an ethylene oxideatmosphere. The pre-sterilized, aseptically packaged tube/bag manifoldsare commercially available (currently from TC Tech; HyClone; St GobainPerformance Plastics, for example) and are used for the manual transferof solutions. Typically, the solution transfer procedure requires atechnician to operate a peristaltic pump and to manually open and closetube clamps for diverting the solution from the reservoir to the storagebags. Although this procedure reduces the cleaning efforts and cleaningvalidation expense, operator interaction and time still are required,and these approaches are dependent upon operator expertise forconsistent accuracy and precision.

Dispensing approaches having automated features (which can includesensors, monitors and programmable controllers) are generally known.Keys et al. U.S. Pat. No. 5,480,063 and U.S. Pat. No. 5,680,960 describefluid dispensing units which control fluid volumes in conjunction with aclosed loop approach, which these patents suggest can avoid the need forventing. The fluid to be dispensed exits the closed loop apparatusthrough a fill tube, as directed by a controller. Such approaches do notaddress the cleaning needs and/or cleaning validation costs andexpenses, were these types of systems to be used in pharmaceutical andbiotech industries for dispensing, directing, combining or separatingbiological or chemical fluids.

Prior systems can incorporate diaphragm valves, which come into directcontact with the process solution, and these valves are a potentialsource of contamination. Thus diaphragm valves require costly cleaningvalidation procedures. In addition, such systems typically lack suitablesensors, especially conductivity sensors.

It has been found that, by proceeding in accordance with the presentinvention, significant cost savings and better performance can berealized in a system which incorporates automated, aseptic manifolds andsensors within the field of technology which embraces pre-sterilized,single-use plastic tubing and containers having at least one collapsibleportion. The components and sensors which contact the biological orchemical fluid are each presterilized and disposable after use.

SUMMARY OF THE INVENTION

The present invention is directed to manifold units which include atleast one sensor and which are presterilized and disposable, making themsingle-use units which are sterilized and packaged so as to be usable“off the shelf” and which thus directly address the problem of tediousand time consuming cleaning and testing at the use site. Multipleembodiments are disclosed. Each includes tubing lengths, at least onesensor and a plurality of single-use storage or collection bags, eachhaving multiple inlet and/or outlet passages which are selectivelyopenable and closeable. The tubing lengths can interact with one or morepinch valves which are operable remotely. Remote operation is automatedby a controller programmed to carry out procedures according to aselected embodiment.

It is a general aspect or object of the present invention to provideimproved single-use manifolds with at least one sensor for automated,aseptic transfer of solutions in bio-processing or chemical processingapplications.

Another aspect or object of the present invention is to provide improvedapparatus and method which combine pinch valve use with disposable,sterilized manifold dispenser units that incorporate at least onedisposable sensor.

Another aspect or object of this invention is to provide improvedapparatus and method which greatly reduce the expenditure of time andresources devoted to cleaning procedures for transfer equipment used inpharmaceutical and biological industries and laboratories wherecontamination of biological and/or chemical fluids cannot be tolerated.

An aspect or object of the present invention is to reduce the need forvalidation procedures for equipment used in separation and purificationof fluids such as in conjunction with the preparation, separation,sensing and dispensing of bio-medical products.

Another aspect or object of this invention is that it handlescleanliness requirements for procedures including a sensing function,such as fluid dispensing, preparative chromatography and tangential flowfiltration while automating operation thereof.

Another aspect or object is to integrate disposable conductivity sensorswith the equipment used in the separation and purification of fluids.

Another aspect or object is to provide the ability to connect disposableconductivity sensors with either a system controller or a userinterface.

These and other objects, aspects, features, improvements and advantagesof the present invention will be clearly understood through aconsideration of the following detailed description.

BRIEF DESCRIPTION OF THE DRAWINGS

In the course of this description, reference will be made to theattached drawings, wherein:

FIG. 1 is a somewhat schematic illustration of a single-use,presterilized system which is especially suitable for solution transferand collection;

FIG. 2 is an illustration of the single-use system of FIG. 1 inoperative association with pinch valves, at least one of which isremotely operable;

FIG. 3 is an illustration of the combination of the features of FIG. 1and FIG. 2, shown with means for use to transfer solution through thesystem;

FIG. 4 is a somewhat schematic illustration of a single-use,presterilized system which is especially suitable for use in automatedpreparative chromatography;

FIG. 5 is an illustration of the single-use system of FIG. 4 inoperative association with pinch valves, at least one of which isremotely operable;

FIG. 6 is an illustration of the combination of features of FIG. 4 andFIG. 5, shown with means for use in transferring solution through thesystem;

FIG. 7 is a somewhat schematic illustration of a single-use,presterilized system which is especially suitable for automatedtangential flow filtration procedures;

FIG. 8 is an illustration of the single-use system of FIG. 7 inoperational association with pinch valves, at least one of which isremotely operable;

FIG. 9 is an illustration of the combination of the features of FIG. 7and FIG. 8, shown with means for use to transfer solution through thesystem;

FIG. 10 is an illustration of the single-use system especially suitablefor solution transfer and collection in operational association with atleast one disposable conductivity sensor;

FIG. 11 is an illustration of the single-use system especially suitablefor use in automated preparative chromatography in operationalassociation with a disposable conductivity sensor;

FIG. 12 is an illustration of the single use system especially suitablefor automated tangential flow filtration procedures in operationalassociation with at least one disposable conductivity sensor;

FIG. 13 is a cutaway view of a disposable conductivity sensor;

FIG. 14 is shows the exterior of a user interface;

FIG. 15 is an exemplarily flowchart of the different screens presentedby the user interface;

FIG. 16 is an exemplarily flowchart of the different screens presentedby the user interface when the user selects to run the recalibrationprogram; and

FIG. 17 is an exemplarily flowchart of the different screens presentedby the user interface when the user selects to view the calibration andproduction information.

DESCRIPTION OF THE PARTICULAR EMBODIMENTS

As required, detailed embodiments of the present invention are disclosedherein; however, it is to be understood that the disclosed embodimentsare merely exemplary of the invention, which may be embodied in variousforms. Therefore, specific details disclosed herein are not to beinterpreted as limiting, but merely as a basis for the claims and as arepresentative basis for teaching one skilled in the art to variouslyemploy the present invention in virtually any appropriate manner.

A system particularly designed for use as an automated, aseptic solutiontransfer system is illustrated in FIGS. 1-3. Fluids processed accordingto this invention are variously referred to herein as biotechnologyfluids, pharmaceutical fluids, chemical fluids, and so forth. These areunderstood to be solutions, liquids, gas-including systems, and thelike. In general, these are referred to herein as biotechnology fluid orfluids.

In the pharmaceutical and biotechnology industries, media preparationdepartments typically prepare the solutions used in a solutionproduction protocol which follows good manufacturing practices. Mediapreparation departments are responsible for maintaining solutionrecipes, preparing and storing buffer solutions and other tasksdemanding consistency and accuracy. For example buffer solutions areprepared in large vats, then pumped through a sterilizing filter, suchas one having a porosity of 0.1μ. Typically such solutions need to befilled into presterilized, single use storage bags for later use. Amedia preparation department may also be responsible for providinginoculating solutions to the operators of a bioreactor. At thecompletion of a bioreactor batch, the reactor broth often is filled intosterile storage bags for later processing.

FIG. 1 shows single-use, presterilized components of the invention.Generally, these disposable components are a manifold and transfertubing assembly and a plurality of bags. A plurality of single-usestorage/collection bags 21, 22, 23 are shown. Each has three tubeconnections. The primary inlet tubing consists of an aseptic connector24 and a manual shut-off clamp 25, each of generally known construction.During solution storage, the aseptic connector is covered with an endcap (not shown) to protect the connector 24 from contamination. Themanual shut-off clamp 25 is closed during solution storage. These areshown on a first tube connection 30.

The second tube connection 26 consists of a short piece of tubingconnected to the bag with a closed manual shut-off clamp. This tubingand clamp arrangement is used to relieve any gas and/or pressurebuild-up inside the bag during the filling operation. The third tubeconnection 27 is identical to the second connection and includes a shortpiece of tubing and a clamp. This can be used as an auxiliary inletand/or outlet for recirculation of the bag contents.

During a typical bag-filling operation, the first and/or last collectionbag can serve the purpose of quality control bags. Often these qualitycontrol bags will be smaller in volume, such as one liter. During theinitial system priming cycle, the first such quality assurance (QA) bagis filled with process solution. At the end of the dispensing cycle whenall of the bags containing the product of the operation, usually largerin volume that the QA bag(s), have been filled, the second QA bag isfilled. The solutions contained in the QA bags are subsequently analyzedfor contamination or for other quality assurance needs.

When the bag-filling process is completed, the manual shut-off clamps oneach bag are closed and the aseptic tube connections are disconnected.During storage, the aseptic connector ends are protected with end caps(not shown).

Turning now to the single-use, sterilized manifold and transfer tubingassembly of FIG. 1, one such unit is generally shown at 28. Thisrepresents a generalized manifold for automated solution transfer. Aninlet end portion 29 of transfer tubing 31 of the unit 28 is forcommunication with a container, such as a vat, of solution, typicallysterile solution. Sterilized manifold and transfer tubing assembly 28 isshown with an optional, in-line pressure sensor 32 and a single-usesterilizing filter 33. An end portion having serially connected endportions are downstream of the illustrated filter 33. By a suitablemovement imparting device, solution moves from the vat or reservoirthrough the sensor 32 (if included) and filter 33 and then is seriallydiverted into the single-use, sterilized storage bags.

FIG. 2 shows a plurality of pinch valves 41, 42, 43 and their respectiverelative positions with respect to the storage bags. Some or all of thevalves can be operated remotely and typically will be pneumatically orelectrically activated. A typical set up will have capacity for up totwelve pneumatically actuated pinch valves or more. A like number ofstorage bags can be accommodated. FIG. 2 shows the relative positions ofthe pinch valves in association with the optional pressure sensor andthe single-use, sterilizing filter. FIG. 3 shows the relative positionof the manifold and transfer tubing assembly 28 with the vat 44 and thepump head of a pump unit 45. Preferably, the pump is a high-accuracy,low-shear peristaltic pump which provides gentle and reproducible bagfilling. An example is a Watson Marlow 620 RE peristaltic pump head.

Access to the storage bags is provided via the pinch valves. The pinchvalves are normally closed, and typical pneumatic pinch valves requirepressurized air (for example 80-100 psi) to open. When such a pinchvalve is pressurized, solution is allowed to enter the storage bag whilethe air in the bag escapes through an integral vent filter. The pinchvalve(s) are pneumatic or electrically operated pinch valves (currentlyavailable from ACRO Associates, Inc). They are installed external to thetubing and are operated by a multi-valve controller (currently availablefrom SciLog Inc.), or another computer-based process logic control (PLC)device. The external pinch valves divert the solution inside themanifold without compromising the sterile environment inside the tubing.Diaphragm valves used in other systems are in constant contact with theprocess solution, whereas pinch valves do not contact the processsolution.

The optional, disposable pressure sensor 32 continuously monitors thefilter back pressure. This sensor can provide information to a suitablecontroller to avoid undesired events. For example, a controller canissue an alarm when a safe, user-defined pressure limit has beenexceeded, indicating that the capacity of the sterilizing filter hasbeen exhausted. Details in this regard are found in U.S. Pat. Nos.5,947,689, 6,350,382 and 6,607,669. These patents and all otherreferences noted herein are incorporated by reference hereinto.

The controller can be a stand-alone unit or be associated with anotherdevice. In a preferred arrangement, the controller is associated withthe pump unit 45. This is shown at 46 in FIG. 3. Whatever form it takes,the controller controls operation of the remotely operable pinchvalve(s). The batch filling rate as well as the batch volume deliveredinto each storage bag is user-programmable via software residing in thecontroller. The controller provides automated bag filling by volume,weight or based on filling time and pump rate.

Typically, a user-determined program will be provided for the automatedfilling of storage bags according to FIGS. 1-3. This is described interms of a SciPro controller of Scilog, Inc., generally described inU.S. Pat. Nos. 5,947,689, 6,350,382 and 6,607,669. With theseapproaches, excessive pressure build-up, as well as associated leaks andbag failures are prevented. For example, when so programmed, thecontroller will stop all pumping action when a user-defined safepressure limit is exceeded.

An exemplary solution transfer program for controlling the manifold isas follows. In a SciPro edit mode, the user enters and stores amulti-bag metering program. The following is an example of a simpleprogram to fill three, 20-liter storage bags 21, 22, 23.

Filling Program Example

000 START The following program steps are entered in an edit mode 001 CWMotor Runs Clockwise 002 RUN Motor is tuned “ON” 003 V 100000 PinchValve 41 is Energized, other pinch valves are De-energized 004 RATE: 5.0l/min Pump Rate 5 liters per minute 005 TIME: 00:04:00 Pump Runs 4minutes, Bag 21 is filled with 20 Liters 006 STOP Pump “Off”, 007 V020000 Pinch Valve 42 is Energized, other valves pinch are De-energized008 TIME: 00:00:02 2 Second Time delay 009 RUN Pump “ON” 010 RATE: 5.0l/min Pump Rate 5 liters per minute 011 TIME: 00:04:00 Pump Runs 4Minutes, Bag 22 is filled with 20 Liters 012 STOP Pump “Off” 013 V003000 Pinch Valve 43 is Energized, other pinch valves are De-energized014 TIME: 00:00:02 2 Second Time Delay 015 RUN Pump “ON” 016 RATE: 5.0l/min Pump Rate 5.0 liters per minute 017 TIME: 00:04:00 Pump Runs 4Minutes, Bag 23 is filled with 20 Liters 018 STOP Pump “Off” 019 V000000 All Pinch Valves are De-energized 020 COUNT: 1 The Program Steps000 to 020 are executed once 021 END

Changes in the RATE and TIME program steps will accommodate any storagebag volume. Additional “RUN” program blocks can be inserted to increasethe number of bags (up to 12 in the example) to be filled. However, ananalogous software program can be generated in which storage bags arefilled based upon either VOLUME or WEIGHT program commands. A scale withan appropriate capacity is required for bag filling by weight. Anoptional scale or load cell 47 can be provided to supply data to thecontroller in this regard. It will be appreciated that this embodimentmeters user-defined volumes of fluid, then automatically switches to thenext empty storage bag to be filled.

A second embodiment, which is generally illustrated in FIGS. 4-6,achieves automated preparative chromatography. In preparativechromatography, process solution containing the bio-molecule of interestis pumped through a column of gel-like particles (stationary phase)suspended in a liquid. The bio-molecule of interest specificallyinteracts (via ion-ion interactions, hydrophobic interactions, sizeexclusion, affinity, for example) with the stationary phase therebyretarding the progress of the bio-molecule through the column. Ideally,other dissolved biomaterials will interact only weakly with thestationary phase and thus will exit the column quickly.

The result is a concentration as well as a separation of thebio-molecule from the rest of the process solution matrix. Theintroduction of an elution buffer will change the local chemicalenvironment of the stationary phase, thereby causing the bio-molecule tobe released and thus able to be collected outside the column in arelatively small volume of elution buffer.

In automated preparative chromatography, the column containing thestationary phase first is washed and/or equilibrated with an appropriatebuffer solution. This wash and/or equilibration cycle is followed by aloading cycle during which the process solution is pumped through thecolumn. The bio-molecule of interest adheres to the stationary phase.The loading cycle can take many hours, depending on the process solutionvolume and pump rate with which the solution is pumped through thecolumn. The loading cycle is followed by a second wash cycle to removeany un-adsorbed biomaterial off the column.

An elution buffer then is introduced to remove the bio-molecule from thecolumn. This removal of the bio-molecule is accomplished either with astep gradient or a linear gradient. After peak collection has beencompleted, the chromatography column is regenerated and re-equilibratedusing appropriate buffer solutions as generally known in the art.

Manifold and transfer tubing assembly 48 represents a generalizedmanifold for automating preparative chromatography procedures. Inoperation, and utilizing the controller system, the exemplarypneumatically controlled pinch valve 51 is pressurized and thus opened,thereby providing access to the wash and/or equilibration buffer bag 54.At a user-definable pump rate, the wash buffer is pumped through adisposable, in-line pressure sensor 55, through a bubble trap (notshown), through the chromatography column 56, and through a detector orUV flow cell 57. On exiting the flow cell, the wash/equilibration bufferis collected in a waste container or bag 58 while pinch valve 49 ispressurized and thus open.

During the loading cycle, pinch valves 51 and 49 are opened/pressurized,while the pinch valves 52, 53 and 59 remain closed. The pump unit 45pumps the process solution through the manifold system 48, the column 56and the flow cell 57 and is collected in the waste container or bag 58.In some chromatography applications, the process solution exiting theflow cell needs to be stored separately in a “process receiving bag”(not shown) for possible re-processing. Another pinch valve (not shown)would provide access to such a “process receiving bag”.

The loading cycle is followed by a wash cycle (valves 51 and 49 areopen/pressurized, all other pinch valves are closed) which carries awayany un-absorbed material from the column to waste. By opening pinchvalves 53 and 49, elution buffer in bag 63 is introduced into the columnand is initially pumped to waste. However, when the signal from the UVdetector 57 exceeds a user-defined value, pinch valve 59 is openedthereby providing access to a peak collection bag 61 while valve 49 isclosed. On the backside of the eluted peak, valve 59 is again closed,while at the same time, valve 49 is opened.

After the material of interest has been collected in bag 61, thechromatographic column 56 requires regeneration and re-equilibration.The column regeneration process is readily automated via access toappropriate buffer solutions (not shown), which are generally as knownin the art. Depending on the underlying chromatographic complexity ofthe application, access to five or six buffer solutions may be required,and these can be provided in their own single-use bags as desired.Similarly, if multiple product peaks are to be collected, additionalpeak collection bag(s) as well as additional pinch valve(s) may have tobe incorporated into manifold and transfer tubing assembly 48.

The single-use, presterilized components of the manifold and transfertubing assembly 48, shown as a feed section, and of a second tube andbag assembly 64 for chromatographed fluid are shown in FIG. 4. Each ofthe single storage/collection bags 54, 62, 63 shown in FIG. 4 has threetube connections. The primary inlet tubing 65 consists of an asepticconnector 66 and a manual shut-off clamp 67. During solution storage,the aseptic connector is covered with an end cap to protect theconnector from contamination. The manual shut-off clamp is closed duringsolution storage.

The second tube and bag assembly 64 consists of a short piece of tubing68 connected to the bag with a closed manual shut-off clamp 69. Thesecond tubing/clamp arrangement 71 is used to relieve any gas and/orpressure build-up inside the bag during the filling operation. The thirdtube connection 72 is identical to the second tubing/clamp arrangement71 and is used as an auxiliary inlet/outlet for recirculation of the bagcontents.

The single-use storage/collection bags 58 and 61 are connected to theremaining tube manifold 72 as shown in FIG. 4 and FIG. 5. FIG. 5 showsthe relative positions of the pinch valves 51, 52, 53, 49 and 59 and theposition of the pressure sensor 55. FIG. 6 shows the insertion of themanifold tubing into the peristaltic pump head 45 as well as connectionsto the chromatography column 56 and the detector 57.

In a typical chromatography application, the single-use storage bags 54(for wash buffer), 62 (for process solution) and 63 (for elution buffer)have been previously filled, for example by using the embodiment of FIG.1-3. When the chromatography run is completed, the manual shut-offclamps on each collection bag 58 (for waste), 61 (for peak collection),and for process receiving (when desired, not shown) are closed, and theaseptic tube connections are disconnected. During storage, the asepticconnector ends are protected with end caps.

Referring further to the SciPro controller programmed for controllingthe manifold arrangement for chromatography, a mode thereof allows entryand storage of a sequence of simple commands, i.e. RUN, RATE, TIME,VOLUME, P LIMIT 1 and Valve States such as V=000000 (all pinch valvesare closed) or V=123456 (all pinch valves are open).

This controller mode is organized in subprogram blocks. The terminatingstatement of a program block can be a “VOLUME”, “TIME”, “P LIMIT D1 (orD2)” or “N LIMIT D1 (or D2)” statement. The statement “P LIMIT D1=5%”reads: “Positive Slope Signal of Detector D1 with a Threshold Value of5% Full Scale (FS)”. See the Chromatography Program Example.

Chromatography Program Example

000 START Start of 1^(st) Wash Cycle 001 CW Clockwise Motor Direction002 RUN Starts Motor 003 RATE 0.25 L/M Pump Rate During Wash Cycle 004 V100050 Wash Buffer 51 Diverted to “Waste” 49 005 VOLUME 1.0 Liters 4Minutes, End of 1^(st) Wash Cycle, TV = 1.0 L 006 RATE 1.00 L/M LoadingRate, Start of Loading Cycle 007 V 020050 Process Solution (52) Divertedto “Waste” (49) 008 TIME: 00:02:00 2 Minutes, End of Loading Cycle, TV =3.0 L 009 RATE 0.25 L/M Start of 2^(nd) Wash Cycle 010 V 100050 WashBuffer (51) Diverted to “Waste” (49) 011 VOLUME 1.0 Liter 4 Minutes, Endof 2^(nd) Wash Cycle TV = 4.0 L 012 V 003050 Elution Buffer (53)Diverted to “Waste” (49) 013 P LIMIT D1 = 5% Threshold Value DetectedStart of Peak Volume Collection 014 V 003400 Elution Buffer (53)Diverted to “Collect” (59) 015 N LIMIT D1 = 10% D1 Threshold Value, Endof Peak Volume Collection 016 V 003050 Elution Buffer (53) Diverted to“Waste” (49) 017 VOLUME 1.0 Liter Elution Volume, End of Elution, TV =5.0 L 018 RATE 0.50 L/M Start of 3^(rd) Wash Cycle 019 V 00050 WashBuffer (51) Diverted to “Waste” (49) 020 TIME 00:02:00 2 Minutes, End of3^(rd) Wash Cycle, TV = 6.0 L 021 STOP Pump Stops, 022 V 000000 AllV-valves Closed 023 END End of Program

For example, in line 014, the SciPro switches from “Waste” to “Collect”when the D1 signal has a positive slope and a value greater than 5% FS(line 013). The statement “N LIMIT D1=10%” reads: “Negative Slope Signalof Detector D1 with a Threshold Value of 10% FS”. In line 016, thecontroller switches from “Collect” to “Waste” when the D1 signal has anegative slope (back side of peak) and a value of 10% FS (line 15).

The user can edit and/or modify the values of: RUN, RATE, TIME, VOLUME,P LIMIT 1, N LIMIT D1 and Valve States at any time during achromatography run. User-designed application programs can be uploadedor downloaded from an external computer at any time by utilizing thecomputer's hyper terminal.

It will be appreciated that, with this embodiment, sequential schedulingof events are achieved. These include sequential scheduling of wash,load and elution cycles. The controller can initiate buffer selection,loading and peak volume collection. Typical in-line concentrationdetectors can be Wedgewood UV and/or pH detectors, which have outputs of4-20 MA outputs which can be monitored simultaneously. A typical pump isa Watson Marlow 620 R peristaltic pump head capable of generating 60 psiat a pump rate of 15 liters per minute.

User-defined detection threshold levels are used for valve switching andpeak volume collection. All solution-handling parameters, such as pumprates, column pressure, and valve positions can be monitored anddocumented in real time and can be printed out or electronicallyarchived.

In a third embodiment, automated tangential flow filtration is carriedout using a modified system designed for this use. Previously referencedU.S. Pat. No. 5,947,689, No. 6,350,382 and No. 6,607,669 disclose theautomation of tangential flow filtration (TFF) procedures. These arecombined with the use of disposable, single-use manifolds, which alsoinclude disposable pressure sensors and single-use, collapsible storagebags and the use of remotely operated pinch valve(s).

A typical TFF application that utilizes a single-use, pre-sterilizedmanifold is shown in FIGS. 7-9. FIG. 7 shows the disposable,pre-sterilized components, including a tubing filtered fluid sectionhaving a permeate collection bag 81 as well as a process solution bag 82within a filtration flow-through section of the tubing. These areaseptically sealed and in a pre-sterilized (for example, irradiated)package. At the beginning of the TFF application, the permeatecollection bag 81 is empty and deflated and has been asepticallyconnected to the TFF manifold. The process solution bag was previouslyfilled, such as by using the system of FIGS. 1-3. The process solutionbag 82 is placed onto an optional scale 83 and connected aseptically tothe rest of the system. In some applications, weight information can beconveyed to the controller in carrying out the control logic.

The pre-sterilized components of this embodiment are shown in FIG. 7.The permeate collection bag 81 has three tube connections. The primaryinlet tubing 84 consists of an aseptic connector 85 and a manualshut-off clamp 86. During solution storage, the aseptic connector iscovered with an end cap to protect the connector from contamination. Themanual shut-off clamp is closed during solution storage.

The second tube connection consists of a short piece of tubing 87connected to the bag with a closed manual shut-off clamp 88. The secondtubing and clamp arrangement is used to relieve any gas and/or pressurebuild-up inside the bag during the filling operation. The third tubeconnection 89 can be identical to the second tubing and clamparrangement and is used as an auxiliary inlet and outlet forrecirculation of bag contents.

Similarly, the process solution bag 82 has three inlet and/or outlettube connections. The first tube connection 91 is used as an outlet topump solution out of the bag. The second tube connection 92 serves as areturn inlet to accommodate the re-circulated retentate. The third tubeconnection 93 again serves to relieve any excessive gas and/or pressurebuild-up inside the bag.

The permeate collection bag and the process solution bag are connectedto the filtration tube manifold, generally designated at 94 in FIG. 7.FIG. 8 shows the relative positions of the pinch valves 95 and 96 andthe position of three pressure sensors 97, 98, 99. FIG. 9 shows theinsertion of the manifold tubing into the head of the peristaltic pumpunit 45.

Prior to starting the pump unit 45, all of the manual shut-off clampsare opened except those clamps that relieve any gas and/or pressurebuild-up inside the bags. Initially the valve 95 is closed and the valve96 is open, while the pump unit 45 starts to recirculate the solutioncontained in the process solution bag 82 through a tangential flowfilter system 101. The air volume contained in the tubing and tangentialflow filter system 101 ends up in the process solution bag 82 where itis vented to the outside through a sterilizing air filter (not shown).Once the optimal pump recirculation rate has stabilized, pinch valve 95is opened and permeate is collected.

The micro filtration or ultra filtration can be carried out either byconstant rate or by constant pressure. Software programs which aresuitable to automate the filtration process through the use of thecontroller 46 are described in U.S. Pat. No. 5,947,689, No. 6,350,382and No. 6,607,669.

The forth, fifth, and sixth embodiments, which are generally illustratedin FIG. 10, FIG. 11, and FIG. 12, respectively, are similar in manyrespects to the first three embodiments illustrated in FIGS. 1-9.However, the systems shown FIGS. 10-12 include at least one conductivitysensor. Any available conductivity sensor may be used with thesesystems, for example, toroidal sensors. The conductivity sensor is apre-sterilized, single-use, disposable, in-line sensor. The embodimentshown in FIG. 13 is a sensor with electrodes.

FIG. 10 shows an aseptic solution transfer system similar to the systemof FIGS. 1-3 and like numbers designate like components. However, inthis embodiment the in-line pressure sensor 32 is replaced with adisposable in-line conductivity sensor 102. During operation, thesolution moves from the vat or reservoir 44 through the sensor 102, thefilter 33, and then is serially diverted into the single use storagebags, 21, 22 and 23. The pinch valves 41, 42, and 43, as describedabove, may be included as desired and may be operated remotely to closethe lines into each storage bag and typically will be pneumatically orelectrically activated.

The conductivity sensor monitors the conductivity levels of thesolution. The levels are reported back either to a user interface, whichdisplays the information, or to the manifold controller 46. Based on theinformation provided by the conductivity sensor or sensors, the manifoldcontroller 46 (or the user interface in some embodiments) may thenmodify the operation of the pump unit 45, open and close the variouspinch valves, start user-determined programs, or stop user-determinedprograms.

The fifth embodiment is generally illustrated in FIG. 11 and is utilizedto achieve automated preparative chromatography. As stated above, inpreparative chromatography, a process solution containing thebio-molecule of interest is pumped through a column of gel likeparticles (stationary phase) suspended in a liquid. The bio-molecule ofinterest interacts with the stationary phase while the otherbio-molecules in the process solution will quickly exit the column. Themanifold and transfer tubing assembly 48 represents the generalizedmanifold system as shown in FIGS. 4-6. Unlike the system shown in FIGS.4-6, the fifth embodiment replaces the in-line pressure sensor 55, thedetector 57, or both with an in-line conductivity sensor 155, 157.

The conductivity sensors monitor the conductivity levels of the solutionentering the chromatography column 56 and the conductivity levels as thesolution leaves the column. The levels are reported back either to auser interface, which displays the information, or to the manifoldcontroller 46. Based on the information provided by the conductivitysensors, the manifold controller 46 (or the user interface in someembodiments) may then modify the operation of the pump unit 45, open andclose the various pinch valves, start user-determined programs, or stopuser-determined programs.

The sixth embodiment, shown in FIG. 12, demonstrates how conductivitysensors may be used in conjunction with a system designed to performautomated tangential flow filtration. The sixth embodiment has the sameoverall configuration as the system shown in FIGS. 7-9, with theaddition of an in-line conductivity sensor 158 which is positioned afterthe pressure sensor 98 and before the pinch valve 96.

The conductivity sensors monitor the conductivity levels of the fluidpassing to the process solution bag 82. The conductivity levels arereported back either to a user interface, which displays theinformation, or to the manifold controller 46. Based on the informationprovided by the conductivity sensors, the manifold controller 46 (or theuser interface in some embodiments) may then modify the operation of thepump unit 45, open and close the various pinch valves, startuser-determined programs, or stop user-determined programs. Theconductivity sensor 158 is useful in TFF as it monitors theconcentration or absence of molecules passing through the tubing to theprocess solution bag 82. For example, if the conductivity sensormeasures abnormally high conductivity levels during the cleaning oroperation of the tangential flow filter, it may signal to the controlleror user that the filter is defective. On the other hand, if theconductivity sensor measures abnormally low conductivity levels duringthe cleaning or operation of the tangential flow filter, it may signalthat the filter or tubing is clogged.

The preferred embodiment of an in-line conductivity sensor has twocomponents: the user interface or the controller 46 and the disposablesensor assembly module. Further description of the in-line, single-useor disposable conductivity sensor is found in U.S. Pat. Nos. 7,788,047,7,857,506 and 7,927,010 and in U.S. Patent Application Publication No.2009/0180513, entitled “Disposable, Pre-Calibrated, Pre-ValidatedSensors for use in Bio-processing Applications,” incorporated hereintoby reference. However, in other embodiments, the functionality of eachcomponent may be combined with or moved to the other component.

The disposable sensor assembly module, generally designated as 200 inFIG. 13, contains inexpensive components. Typically, the sensor assemblymodule contains a short tubular fluid conduit 202 and a sensing portion,generally designated as 201, which includes electrodes 203, a printedcircuit board (PCB) 204 and a sensor-embedded non-volatile memory chip(not-shown). In this embodiment, four electrode pins 203 arepress-fitted through four linearly arranged holes in the fluid conduitwall 202, and are placed in the pathway of fluid progressing through thesystem that is connected at both ends of the fluid conduit 202. Theelectrodes 203 and holes are epoxied, connected or sealed into place toprevent leaks or contamination. The PCB 104 is enclosed in a sheath 205.To prevent contamination and to make the assembly 201 impervious to anyliquid, the combination of the sheath 205, PCB 204, electrodes 203 andto some extent the fluid conduit 202 is sealed in an exterior housing206.

Toroidal conductivity sensors may be used in place of the electrodes inthe sensor assembly 202. The toroids of the toroidal sensors may bearranged in a non-obtrusive manner around the fluid circuit. Typically,two toroids are used. One toroid is used to “drive” or induce a currentthrough the fluid, while the other “senses” or measures the inducedcurrent through the fluid.

The electrodes or toroids are connected to the PCB 204. The PCB maycontain various components, such as a thermistor to measure thetemperature of the fluid in the fluid circuit 202 or a non-volatilememory chip or EEPROM. The PCB is connected to a user interface, controlunit, or controller 46. The controller 46 or user interface connects toand accesses the PCB 204, its components, and the electrodes 203 by aplug-in wires or leads (not shown).

The controller 46 or the user interface produces the current that drivesthe electrodes or toroids and measures the conductivity by measuring thecurrent on the “sensing” electrodes or toroids. The conductivity of thefluid passing through the fluid conduit is measured by driving a currentthrough one or more electrodes, and then using the remaining electrodesto measure the current that passes through the fluid. The current or thevoltage drop measured is proportional to the conductivity of the fluidpassing through the fluid conduit.

The user interface or controller 46 may access calibration informationstored in the non-volatile memory of the sensor. During production ofthe disposable sensors 200, small variations in the design and placementof the electrodes 203 as well as variations in the accuracy of thethermistors may lead to inaccurate conductivity measurements. However,each sensor is individually calibrated to account for the adverseeffects due to these small variations. The sensor specific calibrationinformation is stored in the non-volatile memory of the sensor.

This calibration information may include a temperature offset and aconductivity constant. The temperature offset represents the lineardifference between the known temperature of the fluid and thetemperature measure by the sensor at the time of calibration. Theconductivity constant represents the difference between the knownconductivity of the fluid and the conductivity measure by the sensor atthe time of calibration. When measuring the conductivity of the fluid inthe fluid conduit, the user interface or controller 46 will retrieve thecalibration information to use in the calculations for conductivity. Thetemperature offset and conductivity constant are later utilized by theuser interface or controller 46 to calculate the actual conductivity ofthe biotechnology fluid passing through the fluid conduit 202.

The calibration information may also include information about themethod of calibration, the statistical variance among different sensorsin the same lot, and the date when the sensor was last calibrated.

In addition to the calibration information, production information maybe stored in the non-volatile memory on the sensor. Productioninformation may include items such as the date, time, or lot number forwhen the sensor was manufactured.

FIG. 14 shows a possible embodiment of a user interface, as generallydesignated 210. As stated above, the sensor 200 may be connected toeither the controller 46 or a user interface 210. While both the userinterface 210 and the controller 46 may provide the same functions anddisplay similar information, the embodiment of user interface 210 inFIG. 14 may be advantageous. The user interface 210 is somewhat moreportable in comparison to an entire manifold system or controller 46,may be utilized separately from the entire system, and allows for eitherthe user interface or components of the system to be independentlyupgraded or replaced. Other embodiments might replace the controller 46,with the user interface 210. In these embodiments, the smaller userinterface 210 also has control logic to receive data from the system andto operate the various valves and pumps. In keeping with the invention,the terms user interface and controller may be used interchangeably.

The user interface 210 has a display 211 and several input keys on itsface. These keys include the Menu key 222, the Up key 223, the Down key224, the Re-Cal key 225, the Enter key 226, the Exit key 227 and theSensor On/Sensor Standby key 228. To turn the user interface on, theuser must press the Sensor On key 228. During normal operation, thedisplay 211 typically reports the conductivity of the fluid beingmeasured by the system in Siemens, the temperature of the fluid indegrees Centigrade, the percent of total conductivity, and a graphicalrepresentation of the percent of total conductivity.

The Menu key 222 allows users to progress through different menus asshown in FIG. 15. The display screen 221 initially presents “RUN” screen230, which typically displays the conductivity of the fluid beingmeasured by the system in Siemens, the temperature of the fluid indegrees Centigrade, the percent of total conductivity, and a graphicalrepresentation of the percent of total conductivity. If the userrepeatedly presses the Menu key 222, the screen 221 will display theHigh Conductivity Value 231 (for example 80,000 μS) and then the LowConductivity Value 232 (for example 0 μS). If the user continues topress the Menu Key 222, the user interface 220 will display thecalibration information retrieved from the non-volatile memory of thesensor.

The user interface does not necessarily have to use the calibrationinformation stored on the sensor. In the illustrated embodiment, theuser may modify the calibration information utilized by the userinterface 220 without permanently modifying the information stored inthe non-volatile memory on the sensor. The user may manually change thecalibration information utilized by the user interface 220 by selectingthe Up or Down arrow keys 223, 224 when presented with the correspondingscreen.

The modifiable calibration information may include the ReferenceTemperature 233, the Temperature Coefficient 234, and the TemperatureOffset 235. By pressing the Menu Key 222, the user may modify by usingthe Up or Down arrow keys 223, 224 the units in which conductivity isdisplayed 236, the setting for the serial port 237, the different printtimes for the print option 238, the maximum conductivity measurement atwhich point the user interface 220 produces a high audible alarm 239 orlow audible alarm 240. The user may also select to restore or re-installthe factory calibration values 241, or change the date 242 and time 243.When presented with any of the above mentioned options, the user mayreturn the user interface to normal operations without changing theoption by pressing the Exit Key 227.

The user may also re-calibrate the sensor or overwrite the calibrationinformation stored in the non-volatile memory chip by selecting theRe-cal key 225, which runs the re-calibration program. As shown in FIG.16, the recalibration program displays the calibration information onthe display screen 221. The user can scan through the calibrationinformation by using the Up and Down arrow keys 223, 224. By pressingthe Menu key 222, the user may select a specific piece of calibrationinformation, such as the Pump Low Calculation solution 244, ExternalCalibration Data 245 and 246, and the High Pump Calibration 247, 248,and 249. The user may then modify the value for each piece ofcalibration information by selecting the Up or Down keys 223, 224.

After the information is modified, the new value overwrites the storedinformation in the non-volatile memory of the sensor when the userpresses the Enter Key 225. The display 211 will then report the currentreadings 250 as computed using the new calibration information. In thefuture, when the user selects “Factory Reset” 241, the current settingsof the user interface are replaced with those values entered by the userduring the last recalibration program. However, if the user wants to endthe recalibration program without changing the options, he or she needonly press the Exit key 227.

The user interface 220 may also include a sensor key (not shown). Asshown in FIG. 17, when the user presses the Sensor key, the userinterface retrieves the production information and calibrationinformation stored in the non-volatile memory of the sensor. Thecalibration information may include information that was replaced by therecalibration program. Initially, this operation displays the unique IDnumber for the sensor 251. By pressing the Menu key 222, the user mayview other calibration information 252, such as the type of solutionused during calibration as shown, the temperature of the calibrationsolution, and the statistical information for the sensor. The user mayalso view the date when the sensor was last calibrated or recalibrated253. The user may return the user interface to normal operations 254 bypressing the Exit Key 227.

It will be understood that the embodiments of the present inventionwhich have been described are illustrative of some of the applicationsof the principles of the present invention. Numerous modifications maybe made by those skilled in the art without departing from the truespirit and scope of the invention.

1. A manifold system for biotechnology uses, comprising: a manifold unitwhich is pre-sterilized and disposable so as to be adapted forsingle-time usage, including: (a) at least one length of tubing havingat least one inlet end portion, at least one outlet end portion, anoutside surface, and an inside surface which is sterilized for passageof a biotechnology fluid therethrough, (b) a plurality of single-usebags, each bag having an access port, and (c) at least one asepticconnector that operatively connects said length of tubing with each saidsingle-use bag; a plurality of pinch valves remotely operable to engagesaid outside surface of the length of tubing; and a controller whichcontrols operation of said pinch valves, said controller having controllogic which dictates the timing of opening and closing of said remotelyoperable pinch valves.
 2. The system in accordance with claim 1, whereinsaid control logic of the controller dictates the rate of flow of thebiotechnology fluid.
 3. The system in accordance with claim 1, whereinsaid control logic of the controller determines the extent of filling ofat least one of the single-use bags by processing data monitored by thesystem to achieve filling of the single-use bag by volume, by weight, orby flow rate and filling time.
 4. The system in accordance with claim 2,wherein said control logic of the controller determines the extent offilling of at least one of the single-use bags by processing datamonitored by the system to achieve filling of the single-use bag byvolume, by weight, or by flow rate and filling time.
 5. The system inaccordance with claim 1, wherein said control logic is operable toactivate flow of the biotechnology fluid and opens a first remotelyoperable pinch valve for a length of time needed to flow a selectedvolume or weight of biotechnology fluid into a first single-use bagassociated with the first remotely operable pinch valve, wherein saidcontrol logic is operable to activate flow of the biotechnology fluidand opens a second remotely operable pinch valve for a length of timeneeded to flow a selected volume or weight of biotechnology fluid into asecond single-use bag associated with the second remotely operable pinchvalve, and wherein said control logic is operable to activate flow ofthe biotechnology fluid and opens a further remotely operable pinchvalve for a length of time needed to flow a selected volume or weight ofbiotechnology fluid into a third said single-use bag associated with thethird remotely operable pinch valve until a user-selected number ofsingle-use bags are filled.
 6. The system in accordance with claim 1,wherein said control logic is operable to activate flow of thebiotechnology fluid and opens one of the pinch valves for a length oftime needed to flow a selected volume or weight of biotechnology fluidinto a single-use bag associated with that pinch valve, and wherein saidcontrol logic is operable to activate flow of the biotechnology fluidand opens another of the pinch valves for a length of time needed toflow a selected volume or weight of biotechnology fluid into anotherpinch valve until a user-selected number of single-use bags are filled.7. The system in accordance with claim 1, further including a single-usesterilizing filter positioned along said length of tubing such that thebiotechnology fluid can flow therethrough at a location upstream of saidoutlet and portion.
 8. The system in accordance with claim 1, whereinsaid outlet end portion of the tubing has a plurality of seriallyarranged outlet passageways each having one of said aseptic connectorsfor operable connection with one of said single-use bags, and whereinone of said pinch valves controls passage of the biotechnology fluidfrom the tubing to the single-use bag.
 9. The system in accordance withclaim 8, further including a single-use separation component selectedfrom the group consisting of a separation unit, a purification unit, asterilizing filter and a combination thereof positioned along saidlength of tubing such that the biotechnology fluid can flow therethroughat a location upstream of said outlet end portion.
 10. The system inaccordance with claim 8, further including a disposable pressure sensorpositioned along said length of tubing such that the biotechnology fluidcan flow therethrough at a location upstream of said outlet end portion.11. The system in accordance with claim 9, further including at leastone disposable pressure sensor positioned along said length of tubingsuch that the biotechnology fluid can flow therethrough at a locationselected from the group consisting of upstream, downstream and bothupstream and downstream of said single use separation component andupstream of said outlet end portion.
 12. The system in accordance withclaim 1, wherein said system is for automated preparativechromatography, wherein said tubing is in at least two sectionsincluding a chromatography feed section and a chromatographed fluidsection, wherein said chromatography feed section has an outlet and aplurality of serially arranged inlet passageways each having one of saidaseptic connectors operably connected with one of said single-use bags,wherein said chromatographed fluid section has an inlet and said outletend portion of the tubing has a plurality of serially arranged outletpassageways each having one of said aseptic connectors operablyconnected with said single-use bag.
 13. The system in accordance withclaim 12, further including a disposable pressure sensor positionedalong said tubing chromatography feed section such that thebiotechnology fluid can flow therethrough at a location upstream of saidoutlet end portion.
 14. The system in accordance with claim 12, furtherincluding a chromatography column between said outlet of thechromatography feed section of the tubing and said inlet of thechromatographed fluid section of the tubing.
 15. The system inaccordance with claim 1, wherein said system is for tangential flowfiltration, wherein one said single-use bag is a process solution bagand another said single-use bag is a permeate collection bag, whereinsaid tubing is in at least two sections including a filtrationflow-through section and a filtered fluid section, said filtrationflow-through section includes said process solution bag, said filteredfluid section includes said permeate collection bag, and furtherincluding a disposable filter between said filtration flow-throughsection and said filtered fluid section, whereby fluid from said processsolution bag can be filtered through said disposable filter andcollected in said permeate collection bag.
 16. The system in accordancewith claim 15, wherein said inlet end is within said filtrationflow-through section and in operative communication with said processsolution single-use bag, said filtration flow-through section furtherincludes a recirculation length having one of said pinch valves betweenan exit port of said disposable filter and said process solutionsingle-use bag.
 17. The system in accordance with claim 15, furtherincluding a disposable pressure sensor positioned along said filtrationflow-through section tubing such that the biotechnology fluid can flowtherethrough at a location upstream of said disposable filter.
 18. Thesystem in accordance with claim 16, further including a disposablepressure sensor positioned along said filtration flow-through sectiontubing such that the biotechnology fluid can flow therethrough at alocation downstream of said disposable filter.
 19. The system inaccordance with claim 15, further including a disposable pressure sensorpositioned along said filtered fluid length of tubing such that thebiotechnology fluid can flow therethrough at a location between saiddisposable filter and said permeate collection single-use bag.
 20. Thesystem in accordance with claim 1, wherein said primary access port ofat least one of the single-use bags includes a shut-off clamp, andwherein said single-use bag further includes an access port thatreleases gas or pressure build-up from said bag, an auxiliary accessport, and a shut-off clamp for said access port and for said auxiliaryaccess port.
 21. A manifold system for biotechnology uses, comprising: amanifold unit which is pre-sterilized and disposable so as to be adaptedfor single-time usage, including: (d) at least one length of tubinghaving at least one inlet end portion, at least one outlet end portion,an outside surface, and an inside surface which is sterilized forpassage of a biotechnology fluid therethrough, (e) at least onesingle-use bag having an access port, and (f) at least one asepticconnector that operatively connects said length of tubing with said atleast one single-use bag; a plurality of pinch valves remotely operableto engage said outside surface of the length of tubing; and a controllerwhich controls operation of a plurality of said pinch valves, saidcontroller having control logic which dictates the timing of opening andclosing of a plurality of said remotely operable pinch valves.
 22. Thesystem in accordance with claim 21, wherein said control logic of thecontroller dictates the rate of flow of the biotechnology fluid.
 23. Thesystem in accordance with claim 21, wherein said control logic of thecontroller determines the extent of filling of the single-use bag byprocessing data monitored by the system to achieve filling of thesingle-use bag by volume, by weight, or by flow rate and filling time.24. The system in accordance with claim 22, wherein said control logicof the controller determines the extent of filling of the single-use bagby processing data monitored by the system to achieve filling of thesingle-use bag by volume, by weight, or by flow rate and filling time.25. The system in accordance with claim 21, further including asingle-use sterilizing filter positioned along said length of tubingsuch that the biotechnology fluid can flow therethrough at a locationupstream of said outlet and portion.
 26. The system in accordance withclaim 21, further including a single-use separation component selectedfrom the group consisting of a separation unit, a purification unit, asterilizing filter and a combination thereof positioned along saidlength of tubing such that the biotechnology fluid can flow therethroughat a location upstream of said outlet end portion.
 27. The system inaccordance with claim 21, further including a disposable pressure sensorpositioned along said length of tubing such that the biotechnology fluidcan flow therethrough at a location upstream of said outlet end portion.28. The system in accordance with claim 26, further including adisposable pressure sensor positioned along said length of tubing suchthat the biotechnology fluid can flow therethrough at a locationdownstream of said single-use separation component filter and upstreamof said outlet end portion.
 29. The system in accordance with claim 21,wherein said primary access port of the single-use bag includes ashut-off clamp, and wherein said single-use bag further includes anaccess port that releases gas or pressure build-up from said bag, andsaid bag and further includes an auxiliary access port, furtherincluding a shut-off clamp for said access port and for said auxiliaryaccess port.
 30. A system for biotechnology uses, wherein said system isfor preparative chromatography, comprising: a manifold unit which ispre-sterilized and disposable so as to be adapted for single-time usage,including: (a) at least one length of tubing having at least one inletend portion, at least one outlet end portion, an outside surface, and aninside surface which is sterilized for passage of a biotechnology fluidtherethrough, (b) a plurality of single-use bags, each having an accessport, (c) a plurality of aseptic connectors operatively connecting saidlength of tubing with said single-use bag, (d) said tubing is in atleast two sections including a chromatography feed section and achromatographed fluid section, (e) said chromatography feed section hasan outlet and a plurality of serially arranged inlet passageways eachhaving one of said aseptic connectors for operable connection with saidsingle-use bag, and (f) said chromatographed fluid section has an inlet,and said outlet end portion of the tubing is on the chromatographedfluid section and has a plurality of serially arranged outletpassageways each having one of said aseptic connectors operablyconnected with one of said single-use bags; and at least one pinch valveremotely operable in response to a signal remote from said pinch valve,said pinch valve engaging said outside surface of the length of tubingat a discrete location therealong for that pinch valve and independentlyselectively allowing or stopping flow of the biotechnology fluid throughsaid inside surface of the length of tubing at said discrete locationfor that pinch valve, and wherein said pinch valve controls passage ofthe biotechnology fluid from one of said single-use bags to thechromatography feed section.
 31. The system in accordance with claim 30,further including a disposable pressure sensor positioned along saidtubing such that the biotechnology fluid flows therethrough at alocation upstream of said outlet passageways.
 32. The system inaccordance with claim 30, further including a chromatography columnbetween said outlet of the chromatography feed section of the tubing andsaid inlet of the chromatographed fluid section of the tubing.
 33. Thesystem in accordance with claim 30, further including a single-useseparation component selected from the group consisting of a separationunit, a purification unit, a sterilizing filter and a combinationthereof positioned along said tubing such that the biotechnology fluidflows therethrough at a location upstream of said outlet passageways.34. A system for biotechnology uses, wherein said system is fortangential flow filtration, comprising: a manifold unit which ispre-sterilized and disposable so as to be adapted for single-time usage,including: (a) at least one length of tubing having at least one inletend portion, at least one outlet end portion, an outside surface, and aninside surface which is sterilized for passage of a biotechnology fluidtherethrough, (b) a plurality of single-use bags, each having an accessport, one said single-use bag is a process solution bag and another saidsingle-use bag is a permeate collection bag, (c) said tubing is in atleast two sections including a filtration flow-through section and afiltered fluid section, said filtration flow-through section includessaid process solution bag, said filtered fluid section includes saidpermeate collection bag, (d) an aseptic connector that operativelyconnects said length of tubing with said single-use bag, and (e) adisposable filter between said filtration flow-through section and saidfiltered fluid section, whereby fluid from said process solution bag isfiltered through said disposable filter and is collected in saidpermeate collection bag; and at least one pinch valve remotely operablein response to a signal remote from said pinch valve, said pinch valveengaging said outside surface of the length of tubing at a discretelocation therealong for that pinch valve and independently selectivelyallowing or stopping flow of the biotechnology fluid through said insidesurface of the length of tubing at said discrete location for that pinchvalve.
 35. The system in accordance with claim 34, wherein said inletend is within said filtration flow-through section and in operativecommunication with said process solution single-use bag, and saidfiltration flow-through section further includes a recirculation lengthhaving said pinch valve between an exit port of said disposable filterand said process solution single-use bag.
 36. The system in accordancewith claim 34, further including a disposable pressure sensor positionedalong said tubing such that the biotechnology fluid can flowtherethrough at a location upstream of said disposable filter.
 37. Thesystem in accordance with claim 35, further including a disposablepressure sensor positioned along said recirculation length of tubingsuch that the biotechnology fluid can flow therethrough at a locationbetween said disposable filter and said pinch valve along saidrecirculation length.
 38. The system in accordance with claim 34,further including a disposable pressure sensor positioned along saidfiltered fluid length of tubing such that the biotechnology fluid canflow therethrough at a location between said disposable filter and saidpermeate collection single-use bag.
 39. The system in accordance withclaim 34, further including a single-use separation component selectedfrom the group consisting of a separation unit, a purification unit, asterilizing filter and a combination thereof positioned along saidtubing such that the biotechnology fluid can flow therethrough at alocation upstream of said outlet passageways.
 40. A manifold and flowimparting system for biotechnology uses, wherein said system is forpreparative chromatography, comprising: a manifold unit which ispre-sterilized and disposable so as to be adapted for single-time usage,including: (a) at least one length of tubing having at least one inletend portion, at least one outlet end portion, an outside surface, and aninside surface which is sterilized for passage of a biotechnology fluidtherethrough, (b) a plurality of single-use bags, each having an accessport, (c) a plurality of aseptic connectors that operatively connectsaid length of tubing with said single-use bag access port, (d) saidtubing is in at least two sections including a chromatography feedsection and a chromatographed fluid section, (e) said chromatographyfeed section has an outlet and a plurality of serially arranged inletpassageways having one of said aseptic connectors for operableconnection with said single-use bag access port, and (f) saidchromatographed fluid section has an inlet, and the chromatographedfluid section has said outlet end portion of the tubing which has aplurality of serially arranged outlet passageways having one of saidaseptic connectors operably connected with one of said single-use bags;at least one pinch valve remotely operable in response to a signalremote from said pinch valve, the pinch valve engaging said outsidesurface of the length of tubing at a discrete location therealong forthat pinch valve and independently selectively allowing or stopping flowof the biotechnology fluid through said inside surface of the length oftubing at said discrete location for that pinch valve, wherein the pinchvalve controls passage of the biotechnology fluid from the single-usebag to the chromatography feed section; a chromatography column betweensaid chromatography feed section and said chromatographed fluid section;and a flow imparting unit at a selected location upstream of saidchromatography column.
 41. A manifold and flow imparting system forbiotechnology uses, wherein said system is for tangential flowfiltration, comprising: a manifold unit which is pre-sterilized anddisposable so as to be adapted for single-time usage, including: (a) atleast one length of tubing having at least one inlet end portion, atleast one outlet end portion, an outside surface, and an inside surfacewhich is sterilized for passage of a biotechnology fluid therethrough,(b) a plurality of single-use bags, each having an access port, one saidsingle-use bag is a process solution bag and another said single-use bagis a permeate collection bag, (c) said tubing is in at least twosections including a filtration flow-through section and a filteredfluid section, said filtration flow-through section includes saidprocess solution bag, said filtered fluid section includes said permeatecollection bag, (d) an aseptic connector that operatively connects saidlength of tubing with said single-use bag, and (e) a disposable filterbetween said filtration flow-through section and said filtered fluidsection, whereby fluid from said process solution bag can be filteredthrough said disposable filter and can be collected in said permeatecollection bag; at least one pinch valve remotely operable in responseto a signal remote from said pinch valve, the pinch valve located so asto engage said outside surface of the length of tubing at a discretelocation therealong for that pinch valve and independently selectivelyallowing or stopping flow of the biotechnology fluid through said insidesurface of the length of tubing at said discrete location for that pinchvalve; and a flow imparting unit at a selected location upstream of saiddisposable filter.
 42. An automated manifold and flow imparting systemfor biotechnology uses, wherein said system is for automated preparativechromatography, comprising: a manifold unit which is pre-sterilized anddisposable so as to be adapted for single-time usage, including: (a) atleast one length of tubing having at least one inlet end portion, atleast one outlet end portion, an outside surface, and an inside surfacewhich is sterilized for passage of a biotechnology fluid therethrough,(b) a plurality of single-use bags, each having an access port, (c) aplurality of aseptic connectors that operatively connect said length oftubing with said single-use bag, (d) said tubing is in at least twosections including a chromatography feed section and a chromatographedfluid section, (e) said chromatography feed section has an outlet and aplurality of serially arranged inlet passageways each having one of saidaseptic connectors operably connected with said single-use bag, and (f)said chromatographed fluid section has an inlet, and said outlet endportion of the tubing has a plurality of serially arranged outletpassageways each having one of said aseptic connectors for operableconnection with one of said single-use bags; a plurality of pinchvalves, at least one said pinch valve being remotely operable, eachpinch valve engageable with said outside surface of the length of tubingat a discrete location therealong for that pinch valve and independentlyselectively allowing or stopping flow of the biotechnology fluid throughsaid inside surface of the length of tubing at said discrete locationfor that pinch valve, a first said pinch valve controlling passage ofthe biotechnology fluid from one of the single-use bags to thechromatography feed section, and a second said pinch valve controllingpassage of the biotechnology fluid from the tubing chromatographed fluidsection to the single-use bag of the chromatographed fluid section; achromatography column between said chromatography feed section and saidchromatographed fluid section; a flow imparting unit at a selectedlocation upstream of said chromatography column; and a controller whichcontrols operation of said pump unit and of said remotely operable pinchvalve, said controller having control logic which dictates opening andclosing of said remotely operable pinch valve.
 43. The automated systemin accordance with claim 42, wherein said control logic of thecontroller dictates the rate of flow imparted by said flow impartingunit.
 44. The automated system in accordance with claim 42, wherein saidcontrol logic of the controller determines the extent of filling of thesingle-use bag by processing data monitored by the system to achievefilling of the single-use bag by volume, by weight, or by flow rate andfilling time.
 45. The automated system in accordance with claim 42,wherein said control logic has a loading cycle which activates said flowimparting unit and opens a first and a second remotely operated pinchvalve, the first remotely operable pinch valve is upstream of saidchromatography column and controls egress of process solution from acontainer thereof, the second remotely operable pinch valve isdownstream of said chromatography column and controls access to a firstsingle-use bag; said loading cycle of the control logic precedes anelution cycle which opens a third remotely operated pinch valve which isupstream of said chromatography column and controls egress of elutionsolution from a container thereof and into and through saidchromatography column; and said control logic has a peak valuecollection cycle which activates a fourth remotely operated pinch valvewhich is downstream of said chromatography column and controls access ofsolution into a second single-use bag.
 46. The automated system inaccordance with claim 45, wherein said control logic further includes atleast one wash cycle during which said fourth remotely operated pinchvalve is closed to deny access to said second single-use bag.
 47. Theautomated system in accordance with claim 42, further including adetector downstream of said chromatography column which monitors flowout of said chromatography column for a peak collection value; andwherein said control logic receives peak collection value data from saiddetector for use in said peak value collection cycle.
 48. The automatedsystem in accordance with claim 45, further including a detectordownstream of said chromatography column which monitors flow out of saidchromatography column for a peak collection value; and wherein saidcontrol logic receives peak collection value data from said detector foruse in said peak value collection cycle.
 49. The automated system inaccordance with claim 47, wherein said peak value collection datainclude a threshold value start of peak collection and a threshold valueend of peak collection.
 50. The automated system in accordance withclaim 48, wherein said peak value collection data include a thresholdvalue start of peak collection and a threshold value end of peakcollection.
 51. The automated system in accordance with claim 49,wherein said threshold value start of peak collection is a positiveslope signal, and wherein said threshold value end of peak collection isa negative slope signal.
 52. The automated system in accordance withclaim 50, wherein said threshold value start of peak collection is apositive slope signal, and wherein said threshold value end of peakcollection is a negative slope signal.
 53. An automated manifold andflow imparating system for biotechnology uses, wherein said system isfor tangential flow filtration, comprising: a manifold unit which ispre-sterilized and disposable so as to be adapted for single-time usage,including: (a) at least one length of tubing having at least one inletend portion, at least one outlet end portion, an outside surface, and aninside surface which is sterilized for passage of a biotechnology fluidtherethrough, (b) a plurality of single-use bags, each having an accessport, one said single-use bag is a process solution bag and another saidsingle-use bag is a permeate collection bag, (c) said tubing is in atleast two sections including a filtration flow-through section and afiltered fluid section, said filtration flow-through section includessaid process solution bag, said filtered fluid section includes saidpermeate collection bag, (d) a plurality of aseptic connectors thatoperatively connect said length of tubing with at least one of saidsingle-use bags, and (e) a disposable filter between said filtrationflow-through section and said filtered fluid section, whereby fluid fromsaid process solution bag is filtered through said disposable filter andis collected in said permeate collection bag; at least one pinch valveremotely operable in response to a signal remote from said pinch valve,the pinch valve engageable with said outside surface of the length oftubing at a discrete location therealong for that pinch valve; a flowimparting unit at a selected location upstream of said disposablefilter; and a controller operatively controlling said flow impartingunit and said pinch valve, said controller having control logic whichdictates opening and closing of said remotely operable pinch valve anddictates the rate of flow imparted by said flow imparting unit.
 54. Theautomated system in accordance with claim 53, wherein said control logicof the controller determines the extent of filling of the single-usepermeate collection bag by processing data monitored by the system toachieve filling of the single-use bag by volume, by weight, or by flowrate and filling time.
 55. The automated system in accordance with claim53, further including at least one detector positioned along a locationdownstream of said disposable filter for monitoring a parameter of thefluid within said tubing and for transmitting data on the parameter tothe controller, wherein said control logic receives said data from saiddetector and monitors the flow of fluid through the filtration flowthrough section of the tubing until an optimal recirculation parameteris achieved, at which time said control logic signals that saidfiltration flow through section of the tubing is to be blocked byclosing one of said pinch valves and signals that said filtered fluidsection of the tubing is to be unblocked by opening another of saidpinch valves, whereby filtered fluid begins to flow into said single-usepermeate collection bag.
 56. The automated system in accordance withclaim 55, wherein said detector is a pressure sensor, wherein saidrecirculation parameter is fluid pressure, and wherein said controllogic receives data from said pressure sensor to determine when saidoptimum recirculation pressure is achieved.
 57. The automated system inaccordance with claim 55, wherein said control logic directs the flowimparting unit to modify its flow imparting rate in response to changesin pressure at the pressure sensor so as to maintain a substantiallyconstant selected rate imparted to the fluid by the flow imparting unitand thereby assist in achieving said optimum recirculation pressure. 58.The automated system in accordance with claim 55, wherein said detectoris a fluid flow rate sensor, wherein said recirculation parameter isfluid velocity, and wherein said control logic receives data from saidfluid flow rate sensor to determine when said optimum recirculationfluid velocity is achieved.
 59. The automated system in accordance withclaim 58, wherein said control logic directs the flow imparting unit tomodify its flow imparting rate in response to changes in flow rate atsaid fluid flow rate sensor so as to maintain a substantially constantselected flow rate imparted to the fluid by the flow imparting unit andthereby assist in achieving said optimum recirculation pressure.
 60. Amanifold system for biotechnology uses, comprising: a manifold unitwhich is pre-sterilized and disposable so as to be adapted forsingle-time usage in an automated, aseptic biotechnology solutiontransfer system, including: (g) at least one length of tubing having atleast one inlet end portion, at least one outlet end portion, an outsidesurface, and an inside surface which is sterilized for passage of abiotechnology fluid therethrough, (h) at least one single-use bag havingan access port, and (i) at least one aseptic connector that operativelyconnects said length of tubing with said at least one single-use bag;and at least one pinch valve remotely operable to engage said outsidesurface of the length of tubing.
 61. The system in accordance with claim60, wherein said transfer system further includes a flow imparting unitat a selected location upstream of said pinch valve and a controllerhaving control logic which dictates the timing of opening and closing ofsaid remotely operable pinch valve, and wherein the control logic of thecontroller also dictates the rate of flow imparted by said flowimparting unit.
 62. The system in accordance with claim 61, wherein saidcontrol logic of the controller determines the extent of filling of thesingle-use bag by processing data monitored by the system to achievefilling of the single-use bag by volume, by weight, or by flow impartingrate and filling time.
 63. The system in accordance with claim 61,further including a plurality of single-use bags and a plurality ofpinch valves, wherein said control logic is operable to activate flowimparting action of said flow imparting unit and to open a firstremotely operable pinch valve for a length of time needed to flow aselected volume or weight of biotechnology fluid into a first single-usebag associated with the first remotely operable pinch valve, whereinsaid control logic is operable to activate flowing action of said flowimparting unit and to open a second remotely operable pinch valve for alength of time needed to impart flow of a selected volume or weight ofbiotechnology fluid into a second single-use bag associated with thesecond remotely operable pinch valve, and wherein said control logic isoperable to activate flow imparting action of said flow imparting unitand to open a further remotely operable pinch valve for a length of timeneeded to impart flow of a selected volume or weight of biotechnologyfluid into a third said single-use bag associated with the thirdremotely operable pinch valve until a user-selected number of single-usebags are filled.
 64. The system in accordance with claim 61, furtherincluding a plurality of single-use bags and a plurality of pinchvalves, wherein said control logic is operable to activate flowimparting action of said flow imparting unit and to open one of thepinch valves for a length of time needed to impart flow of a selectedvolume or weight of biotechnology fluid into a single-use bag associatedwith that pinch valve, and wherein said control logic is operable toactivate flow imparting action of said flow imparting unit and to openanother of the pinch valves for a length of time needed to impart flowof a selected volume or weight of biotechnology fluid into another pinchvalve until a user-selected number of single-use bags are filled. 65.The system in accordance with claim 60, further including more than onesaid aseptic connector, more than one said single-use bag and more thanone pinch valve, wherein said outlet end portion of the tubing has aplurality of serially arranged outlet passageways each having one ofsaid aseptic connectors operably connected with one of said single-usebags, and wherein one of said pinch valves controls passage of thebiotechnology fluid from the tubing to the single-use bag.
 66. Thesystem in accordance with claim 60, further including a single-useseparation component selected from the group consisting of a separationunit, a purification unit, a sterilizing filter and a combinationthereof positioned along said length of tubing such that thebiotechnology fluid can flow therethrough at a location upstream of saidoutlet end portion.
 67. The system in accordance with claim 60, furtherincluding a disposable pressure sensor positioned along said length oftubing such that the biotechnology fluid can flow therethrough at alocation selected from the group consisting of upstream of said outletend portion, downstream of said separation unit and upstream of saidoutlet end portion and a combination thereof.
 68. The system inaccordance with claim 60, further including more than one said asepticconnector and more than one said single use bag, wherein said system isfor automated preparative chromatography, wherein said tubing is in atleast two sections including a chromatography feed section and achromatographed fluid section, wherein said chromatography feed sectionhas an outlet and a plurality of serially arranged inlet passagewayseach having one of said aseptic connectors operably connected with oneof said single-use bags, wherein said chromatographed fluid section hasan inlet and said outlet end portion of the tubing has a plurality ofserially arranged outlet passageways each having one of said asepticconnectors operably connected with said single-use bag.
 69. The systemin accordance with claim 68, further including a disposable pressuresensor positioned along said tubing chromatography feed section suchthat the biotechnology fluid can flow therethrough at a locationupstream of said outlet end portion.
 70. The system in accordance withclaim 68, further including a chromatography column between said outletof the chromatography feed section of the tubing and said inlet of thechromatographed fluid section of the tubing.
 71. The system inaccordance with claim 60, wherein said system is for tangential flowfiltration, wherein one said single-use bag is a process solution bagand another said single-use bag is a permeate collection bag, whereinsaid tubing is in at least two sections including a filtrationflow-through section and a filtered fluid section, said filtrationflow-through section includes said process solution bag, said filteredfluid section includes said permeate collection bag, and furtherincluding a disposable filter between said filtration flow-throughsection and said filtered fluid section, whereby fluid from said processsolution bag can be filtered through said disposable filter andcollected in said permeate collection bag.
 72. The system in accordancewith claim 71, wherein said inlet end is within said filtrationflow-through section and in operative communication with said processsolution single-use bag, said filtration flow-through section furtherincludes a recirculation length having one of said pinch valves betweenan exit port of said disposable filter and said process solutionsingle-use bag.
 73. The system in accordance with claim 71, furtherincluding a disposable pressure sensor positioned along said filtrationflow-through section tubing such that the biotechnology fluid can flowtherethrough at a location selected from the group consisting ofupstream of said disposable filter, downstream of said disposablefilter, between said disposable filter and said permeate collectionsingle-use bag and a combination thereof.
 74. The system in accordancewith claim 60, wherein said primary access port of the single-use bagincludes a shut-off clamp, and wherein said single-use bag furtherincludes an access port that releases gas or pressure build-up from saidbag, further including an auxiliary access port and a shut-off clamp forsaid access port and for said auxiliary access port.